Objective To prepare complex of lentivirus-mediated fibroblast growth factor 2(FGF-2)gene which was used to transfect into bone marrow stromal stem cells(MSCs)and then were co-cultured with xenogeneic antigenic cancel-lous bone(XACB),and to observe its therapeutic effect on steroid-induced avascular necrosis of femoral head and its effect on the expression of TNF-αin femoral head tissues.Methods Rabbit MSCs were isolated and cultured in vitro.The lenti-virus-mediated FGF-2 gene was used to transfect the MSCs and then were cultured with XACB to obtain tissue engineering bone of XACB/MSCs/Lv-GFP-FGF-2 complex.Sixty rabbit models of steroid-induced avascular necrosis of femoral head which were established by injection of endotoxin and methylprednisolone were randomly divided into 5 groups with 12 in each group.Core decompression and bone grafting were performed.In group A, bone grafting was not performed,autolo-gous cancellous bone was implanted in the group B, XACB/MSCs were implanted in the group C, XACB/MSCs/Lv-GFP was implanted in the group D,and XACB/MSCs/Lv-GFP-FGF-2 was implanted in the group E.Four rats were executed at 3,6,12 weeks after operation in each group,then we observed the general appearance of femoral head.The positive ex-pression rate of TNF-αwas detected by immunohistochemistry.The real-time PCR was used to detect the expression of TNF-αmRNA in the femoral head tissues.Results Three weeks after operation, a small amount of new bone formation were found in the group E,there were no new bone formation in the other groups.At 6 weeks after operation,the defect ar-eas in the group A were filled with granulation tissue and there were no new bones.The defect areas in the group B,C and D were still visible and the new bones were visible at the edge.Most of the defect areas in the group E were filled with new bones,and bone graft was basically absorbed.At 12 weeks after operation,only a small amount of new bone was formed in the group A,and defect areas of the groups B,C,and D were not repaired completely.The defect in the group E was re-stored to normal cancellous bone.At 3,6,and 12 weeks after operation,the positive expression rate of TNF-αand relative expression of mRNA in the femoral head tissues of the groups B,C,D,and E were decreased gradually(all P<0.05). At 3 weeks after operation,the positive expression of TNF-αand the mRNA relative expression of femoral head in the group E were lower than those in the groups A,B,C,and D(all P<0.05).At 6,12 weeks after surgery,the TNF-αpositive expression rate and mRNA relative expression level of the femoral bone tissue at the same time point were in the following order:group A>group B >group C, group D >group E(all P <0.05).Conclusion The tissue engineering bone XACB/MSCs/Lv-GFP-FGF-2 complex is successfully prepared,and it has a good effect on the treatment of steroid-induced avascular necrosis of the femoral head in rabbits by inhibiting the expression of TNF -αin the femoral head tissues.%目的 制备慢病毒介导成纤维细胞生长因子2(FGF-2)基因转染骨髓基质干细胞(MSCs)与异种脱抗原松质骨(XACB)的组织工程骨,探讨其对兔激素性股骨头缺血坏死的治疗作用及其机制.方法 体外分离培养兔MSCs,利用慢病毒介导FGF-2基因转染MSCs并获得稳转细胞株,将其与XACB进行复合培养,获得组织工程骨XACB/MSCs/Lv-GFP-FGF-2.采用内毒素与甲基强的松龙制备兔激素性股骨头缺血坏死模型60只,随机分为A~E组各12只,各组进行髓芯减压,A 组不进行植骨,B 组植入自体松质骨,C 组植入 XACB/MSCs,D 组植入XACB/MSCs/Lv-GFP,E组植入XACB/MSCs/Lv-GFP-FGF-2.术后3、6、12周各组随机处死4只,观察股骨头大体形态,采用免疫组化法检测股骨头组织TNF-α阳性表达率,Real-time PCR法检测股骨头组织TNF-αmRNA表达.结果 术后3周,E组可见少量新骨形成,其余各组均未见新骨形成.术后6周,A组缺损区由肉芽组织填充,仍无明显新骨形成;B、C、D组缺损区仍清晰可见,边缘可见新骨形成;E组大部分缺损区已被新骨填充,移植骨已基本被吸收.术后12周,A组仅有少量新骨形成,B、C、D组均未完全修复,E组缺损区修复接近正常松质骨.术后3、6、12周,B、C、D、E组股骨头组织TNF-α阳性表达率及mRNA相对表达量均逐渐降低(P均<0.05).术后3周,E组股骨头组织TNF-α阳性表达率及mRNA相对表达量均低于A、B、C、D组(P均<0.05);术后6、12周相同时间点,股骨头组织TNF-α阳性表达率及mRNA相对表达量A组>B组>C、D组>E组(P均<0.05).结论 成功制备了组织工程骨XACB/MSCs/Lv-GFP-FGF-2;其治疗兔激素性股骨头缺血坏死的效果较好,作用机制可能为抑制股骨头组织TNF-α表达.
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